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Research Article

Modifications to a LATE MERISTEM IDENTITY1 gene are responsible for the major leaf shapes of Upland cotton (Gossypium hirsutum L.)

Ryan J. Andres, Viktoriya Coneva, Margaret H. Frank, John R. Tuttle, Luis Fernando Samayoa, Sang-Won Han, Baljinder Kaur, Linglong Zhu, Hui Fang, Daryl T. Bowman, Marcela Rojas-Pierce, Candace H. Haigler, Don C. Jones, James B. Holland, Daniel H. Chitwood, and Vasu Kuraparthy
  1. aDepartment of Crop and Soil Sciences, North Carolina State University, Raleigh, NC 27695-7620;
  2. bDonald Danforth Plant Science Center, St. Louis, MO 63132;
  3. cPlant Science Research Unit, United States Department of Agriculture-Agricultural Research Service, Raleigh, NC 27695-7620;
  4. dDepartment of Plant and Microbial Biology, North Carolina State University, Raleigh, NC 27695;
  5. eCotton Incorporated, Cary, NC 27513

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PNAS January 3, 2017 114 (1) E57-E66; first published December 20, 2016; https://doi.org/10.1073/pnas.1613593114
Ryan J. Andres
aDepartment of Crop and Soil Sciences, North Carolina State University, Raleigh, NC 27695-7620;
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Viktoriya Coneva
bDonald Danforth Plant Science Center, St. Louis, MO 63132;
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Margaret H. Frank
bDonald Danforth Plant Science Center, St. Louis, MO 63132;
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John R. Tuttle
aDepartment of Crop and Soil Sciences, North Carolina State University, Raleigh, NC 27695-7620;
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Luis Fernando Samayoa
aDepartment of Crop and Soil Sciences, North Carolina State University, Raleigh, NC 27695-7620;
cPlant Science Research Unit, United States Department of Agriculture-Agricultural Research Service, Raleigh, NC 27695-7620;
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Sang-Won Han
dDepartment of Plant and Microbial Biology, North Carolina State University, Raleigh, NC 27695;
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Baljinder Kaur
aDepartment of Crop and Soil Sciences, North Carolina State University, Raleigh, NC 27695-7620;
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Linglong Zhu
aDepartment of Crop and Soil Sciences, North Carolina State University, Raleigh, NC 27695-7620;
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Hui Fang
aDepartment of Crop and Soil Sciences, North Carolina State University, Raleigh, NC 27695-7620;
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Daryl T. Bowman
aDepartment of Crop and Soil Sciences, North Carolina State University, Raleigh, NC 27695-7620;
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Marcela Rojas-Pierce
dDepartment of Plant and Microbial Biology, North Carolina State University, Raleigh, NC 27695;
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Candace H. Haigler
aDepartment of Crop and Soil Sciences, North Carolina State University, Raleigh, NC 27695-7620;
dDepartment of Plant and Microbial Biology, North Carolina State University, Raleigh, NC 27695;
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Don C. Jones
eCotton Incorporated, Cary, NC 27513
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James B. Holland
aDepartment of Crop and Soil Sciences, North Carolina State University, Raleigh, NC 27695-7620;
cPlant Science Research Unit, United States Department of Agriculture-Agricultural Research Service, Raleigh, NC 27695-7620;
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Daniel H. Chitwood
bDonald Danforth Plant Science Center, St. Louis, MO 63132;
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  • For correspondence: vasu_kuraparthy@ncsu.edu dchitwood@danforthcenter.org
Vasu Kuraparthy
aDepartment of Crop and Soil Sciences, North Carolina State University, Raleigh, NC 27695-7620;
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  • For correspondence: vasu_kuraparthy@ncsu.edu dchitwood@danforthcenter.org
  1. Edited by Sarah Hake, University of California, Berkeley, CA, and approved November 17, 2016 (received for review August 19, 2016)

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Significance

Leaves are the primary source of photoassimilate in crop plants. A precise understanding of the genetic architecture underlying leaf morphology is critical to engineering climate-resilient crop varieties. An ideal cotton cultivar would produce a lower canopy of broad, normal leaves before transitioning to an upper canopy of highly lobed, okra leaves. Here we show that the major leaf shapes of cotton are controlled by the okra locus, which encodes an HD-Zip transcription factor Gossypium hirsutum LATE MERISTEM IDENTITY1-D1b (GhLMI1-D1b). Using gene silencing, we temporarily induced normal leaf formation in okra, thus validating the candidate gene and creating the leaf shape ideotype in cotton. This study, identifying a single locus responsible for cotton leaf shape, expands the genetic toolbox for breeders to produce superior cotton varieties.

Abstract

Leaf shape varies spectacularly among plants. Leaves are the primary source of photoassimilate in crop plants, and understanding the genetic basis of variation in leaf morphology is critical to improving agricultural productivity. Leaf shape played a unique role in cotton improvement, as breeders have selected for entire and lobed leaf morphs resulting from a single locus, okra (l-D1), which is responsible for the major leaf shapes in cotton. The l-D1 locus is not only of agricultural importance in cotton, but through pioneering chimeric and morphometric studies, it has contributed to fundamental knowledge about leaf development. Here we show that an HD-Zip transcription factor homologous to the LATE MERISTEM IDENTITY1 (LMI1) gene of Arabidopsis is the causal gene underlying the l-D1 locus. The classical okra leaf shape allele has a 133-bp tandem duplication in the promoter, correlated with elevated expression, whereas an 8-bp deletion in the third exon of the presumed wild-type normal allele causes a frame-shifted and truncated coding sequence. Our results indicate that subokra is the ancestral leaf shape of tetraploid cotton that gave rise to the okra allele and that normal is a derived mutant allele that came to predominate and define the leaf shape of cultivated cotton. Virus-induced gene silencing (VIGS) of the LMI1-like gene in an okra variety was sufficient to induce normal leaf formation. The developmental changes in leaves conferred by this gene are associated with a photosynthetic transcriptomic signature, substantiating its use by breeders to produce a superior cotton ideotype.

  • cotton
  • leaf shape
  • okra
  • gene cloning

Footnotes

  • ↵1R.J.A., D.H.C., and V.K. contributed equally to this work.

  • ↵2To whom correspondence may be addressed. Email: vasu_kuraparthy{at}ncsu.edu or dchitwood{at}danforthcenter.org.
  • Author contributions: V.K. designed research; R.J.A., V.C., M.H.F., J.R.T., S.-W.H., B.K., L.Z., H.F., D.T.B., M.R.-P., D.H.C., and V.K. performed research; C.H.H. and D.C.J. contributed new reagents/analytic tools; R.J.A., V.C., M.H.F., J.R.T., L.F.S., M.R.-P., D.C.J., J.B.H., D.H.C., and V.K. analyzed data; and R.J.A., V.C., M.H.F., C.H.H., J.B.H., D.H.C., and V.K. wrote the paper.

  • The authors declare no conflict of interest.

  • This article is a PNAS Direct Submission.

  • Data deposition: The sequences reported in this paper have been deposited in the GenBank database (accession nos. KY293601 to KY293643).

  • This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1613593114/-/DCSupplemental.

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Changes to LMI1 gene alter leaf shape in cotton
Ryan J. Andres, Viktoriya Coneva, Margaret H. Frank, John R. Tuttle, Luis Fernando Samayoa, Sang-Won Han, Baljinder Kaur, Linglong Zhu, Hui Fang, Daryl T. Bowman, Marcela Rojas-Pierce, Candace H. Haigler, Don C. Jones, James B. Holland, Daniel H. Chitwood, Vasu Kuraparthy
Proceedings of the National Academy of Sciences Jan 2017, 114 (1) E57-E66; DOI: 10.1073/pnas.1613593114

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Changes to LMI1 gene alter leaf shape in cotton
Ryan J. Andres, Viktoriya Coneva, Margaret H. Frank, John R. Tuttle, Luis Fernando Samayoa, Sang-Won Han, Baljinder Kaur, Linglong Zhu, Hui Fang, Daryl T. Bowman, Marcela Rojas-Pierce, Candace H. Haigler, Don C. Jones, James B. Holland, Daniel H. Chitwood, Vasu Kuraparthy
Proceedings of the National Academy of Sciences Jan 2017, 114 (1) E57-E66; DOI: 10.1073/pnas.1613593114
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